Search

The increasing number of diversified small-scale farms (DSSF) that raise outdoor-based livestock in the USA reflects growing consumer demand for sustainably produced food. Diversified farms are small scale and raise a combination of multiple livestock species and numerous produce varieties. This 2015-2016 cross-sectional study aimed to describe the unique characteristics of DSSF in California, estimate the prevalence of Shiga toxin-producing

Recent foodborne illness outbreaks have heightened pressures on growers to deter wildlife from farms, jeopardizing conservation efforts. However, it remains unclear which species, particularly birds, pose the greatest risk to food safety. Using11,000 pathogen tests and 1565 bird surveys covering 139 bird species from across the western United States, we examined the importance of 11 traits in mediating wild bird risk to food safety. We tested whether traits associated with pathogen exposure (e.g., habitat associations, movement, and foraging strategy) and pace-of-life (clutch size and generation length) mediated foodborne pathogen prevalence and proclivities to enter farm fields and defecate on crops. Campylobacter spp. were the most prevalent enteric pathogen (8.0%), while Salmonella and Shiga-toxin producing Escherichia coli (STEC) were rare (0.46% and 0.22% prevalence, respectively). We found that several traits related to pathogen exposure predicted pathogen prevalence. Specifically, Campylobacter and STEC-associated virulence genes were more often detected in species associated with cattle feedlots and bird feeders, respectively. Campylobacter was also more prevalent in species that consumed plants and had longer generation lengths. We found that species associated with feedlots were more likely to enter fields and defecate on crops. Our results indicated that canopy-foraging insectivores were less likely to deposit foodborne pathogens on crops, suggesting growers may be able to promote pest-eating birds and birds of conservation concern (e.g., via nest boxes) without necessarily compromising food safety. As such, promoting insectivorous birds may represent a win-win-win for bird conservation, crop production, and food safety. Collectively, our results suggest that separating crop production from livestock farming may be the best way to lower food safety risks from birds. More broadly, our trait-based framework suggests a path forward for co-managing wildlife conservation and food safety risks in farmlands by providing a strategy for holistically evaluating the food safety risks of wild animals, including under-studied species.

International audience; Flavobacterium psychrophilum affects salmonid health worldwide and causes economic losses. The genetic diversity of the pathogen must be considered to develop control methods. However, previous studies have reported both high and low levels of genetic diversity. The present longitudinal study aimed at assessing the genetic diversity of F. psychrophilum at a small temporal and geographic scale. Four farms located on the same watershed in France were studied. Rainbow trout (Oncorhynchus mykiss) batches were monitored, and apparently healthy individuals were sampled over 1 year. A total of 288 isolates were recovered from fish organs (gills and spleen) and eggs. Pulsed field gel electrophoresis revealed high genetic diversity. Multilocus sequence typing performed on a selection of 31 isolates provided congruent results, as follows: 18 sequence types (STs) were found, of which 13 were novel. The mean gene diversity (H = 0.8413) was much higher than that previously reported for this host species, although the sampling was restricted to a single watershed and 1 year. Seven isolates out of 31 were assigned to clonal complex ST10 (CC-ST10), which is the predominant clonal complex in the main salmonid production areas. A split decomposition tree reflected a panmictic population. This finding is important for aquaculture veterinarians in their diagnostic procedure, as the choice of adequate antibiotic treatment is conditioned by the correct identification of the causative agent. Furthermore, this study expands our knowledge on genetic diversity required for the development of an effective vaccine against F. psychrophilum.IMPORTANCE The bacterium Flavobacterium psychrophilum is a serious pathogen in many fish species, especially salmonids, that is responsible for considerable economic losses worldwide. In order to treat infections and to develop vaccines, the genetic diversity of this bacterium needs to be known. We assessed the genetic diversity of F. psychrophilum isolates from apparently healthy rainbow trout raised in several fish farms in the same watershed in France. Two different genotyping methods revealed high diversity. The majority of isolates were unrelated to clonal complex sequence type 10 (CC-ST10), the clonal complex that is predominant worldwide and associated with disease in rainbow trout. In addition, we found 13 novel sequence types. These results suggest that a diverse subpopulation of F. psychrophilum may be harbored by rainbow trout.

The shedding dynamics of foodborne pathogens by wild birds on farmland are not well characterized. This yearlong study sampled wild birds for foodborne pathogens within agricultural lands in northern California. There was a low prevalence of Salmonella spp., Escherichia coli O157:H7, and non-O157 Shiga-toxin producing E. coli (prevalence, 0.34% to 0.50%) identified in bird populations in this study. However, pathogens of public health importance (such as Salmonella Newport, E. coli O157:H7, and STEC O103 and O26) were identified in fecal samples, and two birds carried STEC on their feet or feathers. Identical pathogen strains were shared episodically among birds and between wild geese and free-range cattle. This result suggests a common source of contamination in the environment and potential transmission between species. These findings can be used to assess the risk posed by bird intrusions in produce fields and enhance policy decisions toward the comanagement of food safety and farmland habitat conservation., Current California agricultural practices strive to comanage food safety and habitat conservation on farmland. However, the ecology of foodborne pathogens in wild bird populations, especially those avian species residing in proximity to fresh produce production fields, is not fully understood. In this repeated cross-sectional study, avifauna within agricultural lands in California were sampled over 1 year. Feces, oral swabs, and foot/feather swabs were cultured for zoonotic Salmonella spp., Escherichia coli O157:H7, and non-O157 Shiga toxin-producing E. coli (STEC) and characterized by serotyping and pulsed-field gel electrophoresis. Of 60 avian species sampled, 8 species (13.3%, bird groups of sparrows, icterids, geese, wrens, and kinglets) were positive for at least one of these foodborne pathogens. At the individual bird level, the detection of foodborne pathogens was infrequent in feces (n = 583; 0.5% Salmonella, 0.34% E. coli O157:H7, and 0.5% non-O157 STEC) and in feet/feathers (n = 401; 0.5% non-O157 STEC), and it was absent from oral swabs (n = 353). Several subtypes of public health importance were identified, including Salmonella enterica serotype Newport, E. coli O157:H7, and STEC serogroups O103 and O26. In late summer and autumn, the same STEC subtype was episodically found in several individuals of the same and different avian species, suggesting a common source of contamination in the environment. Sympatric free-range cattle shared subtypes of STEC O26 and O163 with wild geese. A limited rate of positive detection in wild birds provides insights into broad risk profile for contamination considerations but cannot preclude or predict risk on an individual farm. IMPORTANCE The shedding dynamics of foodborne pathogens by wild birds on farmland are not well characterized. This yearlong study sampled wild birds for foodborne pathogens within agricultural lands in northern California. There was a low prevalence of Salmonella spp., Escherichia coli O157:H7, and non-O157 Shiga-toxin producing E. coli (prevalence, 0.34% to 0.50%) identified in bird populations in this study. However, pathogens of public health importance (such as Salmonella Newport, E. coli O157:H7, and STEC O103 and O26) were identified in fecal samples, and two birds carried STEC on their feet or feathers. Identical pathogen strains were shared episodically among birds and between wild geese and free-range cattle. This result suggests a common source of contamination in the environment and potential transmission between species. These findings can be used to assess the risk posed by bird intrusions in produce fields and enhance policy decisions toward the comanagement of food safety and farmland habitat conservation.

Mixed crop-livestock farms (MCLF) integrate livestock and crops using their animals to graze crop residues and/or cover crops. MCLF are considered sustainable because grazing and the manure deposited by livestock enhance soil fertility and recycles farm nutrients. However, livestock manure may introduce enteric foodborne pathogens to the soil, which could contaminate fresh produce. Organic farmers in the United States follow the USDA National Organic Program (NOP) standards, which require 90 or 120 days between incorporating raw manure into the soil and harvest. Although not specifically addressed in NOP, organic farmers using grazing within production fields may also use this standard. The objectives of this study were to generate preharvest data to assess the die-off of generic Escherichia coli (E. coli) in the soil, after cover crops were grazed by sheep; and assess the genetic relatedness of generic E. coli isolates between soil and sheep faecal samples. We conducted a repeated observational study to evaluate the persistence of generic E. coli, as an indicator of faecal contamination and surrogate for STEC, in the soil of two fields (A and B) on an organic MCLF. Results showed a 3.70 log10 reduction in mean generic E. coli concentration MPN in the soil of field A from the highest of 3.70 log10 MPN/g on 48 day postsheep grazing (DPS) to -0.70 log10 MPN/g on 139 DPS. Field B showed a 3.51 log10 reduction in mean generic E. coli concentration in the soil from the highest mean of 3.51 log10 MPN/g on 14 DPS to the lowest mean -0.35 log10 MPN/g on 112 DPS. STEC prevalence in the sheep flock was 4.17% (1/24). Closely related generic E. coli strains were found between soil and faecal samples. Developing research-based waiting periods between grazing and harvest is important to inform best practices for farmers and food safety regulators.

Body condition (i.e., the amount of the energy stored in organs and tissues) is a key parameter that has been related with health, reproductive performance and density of wild ungulates including the wild boar (Sus scrofa). In this wild pig, a reference method to assess body condition has not yet been agreed and different procedures have been used in recent literature. The aim of this work was to generate an easy and reliable method based on biometrical measurements and with the ability to predict body fat in live or die boars. For this, a total of 207 hunted wild boar from three Spanish populations with contrasting food availability were included in this study. Sex, age, biometrical parameters (body weight, total length and chest girth) and body condition indicators (brisket and rump fat thickness, kidney fat index (KFI), ratio between chest girth-total length and scaled mass index) were assessed for each animal. A boosted regression trees (BRT) approach was carried out to find models based on age, sex and biometrical features that predicted brisket fat thickness in the studied animals. BRT models including sex, body weight, total length and chest girth as explanatory variables were able to predict brisket fat thickness in wild boar (68–73% of deviance explained). These models were not influenced by the location of sampling and their predictive values showed a good agreement with real brisket fat thickness (94.1–95.6). Predictive values obtained in BRT models from each area also agreed with food availability suggesting this is a valid indicator of body condition of wild boar in a broad range of environmental conditions.

International audience; Mycobacterium avium ssp. paratuberculosis (MAP) is the etiological agent of paratuberculosis, a disease that affects ruminants worldwide. Despite global interest in the control of this disease, gaps exist in our knowledge of fecal shedding patterns and concurrent serological patterns. This longitudinal study in dairy cattle herds with high MAP seroprevalence in France aimed at accurately describing fecal shedding patterns over 1 year; relating those shedding patterns to individual animal characteristics (age, breed, parity); and exploring the association between fecal shedding patterns and serological patterns. To describe temporal fecal shedding patterns and continuity of shedding, along with the standard quantitative PCR (qPCR) threshold cycle we used a cutoff value that related to low or nonculturable fecal shedding. We also defined a threshold cycle indicative of shedding in high quantities to describe infection progression patterns. Twenty-one herds completed the study, and 782 cows were tested 4 times each. We obtained 4 sets of paired fecal qPCR and serum ELISA results from 757 cows. Although we targeted highly likely infectious animals, we found a large diversity of shedding patterns, as well as high variability between herds in the proportion of animals showing a given pattern. The fecal qPCR results of almost 20% of the final study sample were positioned at least once in the range that indicated low or nonculturable fecal shedding (between the adjusted and the standard cutoff value). Although these animals would typically be classified as non-shedders, they could be important to infection dynamics on the farm. Animals that shed at least twice consecutively and animals that shed in high quantities rarely reverted to negativity. Repeated fecal qPCR can be used to detect temporal fecal shedding traits, and the decision to cull an animal could practically be based on temporal, semiquantitative results. Overall, we found a mismatch between fecal shedding and ELISA seropositivity (637 animals were ELISA-negative 4 times, but only 13% of those animals were qPCR-negative 4 times). We found that having more than 2 ELISA-positive samples was strongly related to persistent and continuous shedding. We suggest that although serological testing is much less sensitive than qPCR, it can also be used, particularly over the course of multiple testing events, to identify animals that are most likely to contribute to the contamination of the farm environment

Diversified farms are operations that raise a variety of crops and/or multiple species of livestock, with the goal of utilising the products of one for the growth of the other, thus fostering a sustainable cycle. This type of farming reflects consumers' increasing demand for sustainably produced, naturally raised or pasture-raised animal products that are commonly produced on diversified farms. The specific objectives of this study were to characterise diversified small-scale farms (DSSF) in California, estimate the prevalence ofSalmonella entericaandCampylobacterspp. in livestock and poultry, and evaluate the association between farm- and sample-level risk factors and the prevalence ofCampylobacterspp. on DSSF in California using a multilevel logistic model. Most participating farms were organic and raised more than one animal species. OverallSalmonellaprevalence was 1.19% (95% confidence interval (CI95) 0.6–2), and overallCampylobacterspp. prevalence was 10.8% (CI95= 9–12.9). Significant risk factors associated withCampylobacterspp. were farm size (odds ratio (OR)10–50 acres: less than 10 acres= 6, CI95= 2.11–29.8), ownership of swine (OR = 9.3, CI95= 3.4–38.8) and season (ORSpring: Coastal summer= 3.5, CI95= 1.1–10.9; ORWinter: Coastal summer= 3.23, CI95= 1.4–7.4). As the number of DSSF continues to grow, evaluating risk factors and management practices that are unique to these operations will help identify risk mitigation strategies and develop outreach materials to improve the food safety of animal and vegetable products produced on DSSF.

Author(s): Rivadeneira, Paula; Kross, Sara; Navarro-Gonzalez, Nora; Jay-Russell, Michele | Abstract: Animal intrusion causes significant agricultural losses each year. Wild and domestic animals destroy crops by eating and trampling them, and can pose food safety risks due to the deposition of feces on or near the crops. Birds are one of the most challenging animals to keep out of agricultural fields. Growers try countless methods to deter them, including visual, auditory, tactile, and olfactory means. While some of these methods work some of the time, none provide stand-alone protection all the time. Recently there has been interest in developing technology-based solutions to deter nuisance birds in agricultural settings, while others are exploring more natural methods, including falconry. We provide a general overview of bird deterrent methods that are currently in use in agricultural settings, and explore options for novel methods. We found that very few independent scientific studies have been conducted to assess the efficacy of most bird deterrent methods. Ultimately, a multi-tiered approach using integrated pest management techniques will likely be most useful as it can be tailored to meet the needs of individual farmers.

International audience; The Western European population of wild boar (Sus scrofa) has increased its distribution over the past several decades, and some populations have colonized areas strongly influenced by human activity. Wild boars are known carriers of antibiotic-resistant bacteria acquired from the environment, and urban populations of wild boars may be more exposed than their rural counterparts. In this work, we compared the frequency of antibiotic resistance in indicator bacteria (Escherichia coli, Enterococcus faecalis, Enterococcus faecium) isolated from urban wild boars with that from rural wild boars in NE Spain. We further assessed whether bacterial isolates from the urban wild boars had a higher probability of showing antibiotic resistance when their host was highly associated to urban features. Seventy-two and 100 bacterial isolates from urban and rural habitat, respectively, were screened for antibiotic resistance against a set of antibiotics (13 per bacterial species). We found a significantly higher frequency of E. faecium showing resistance to tetracycline (70.0% vs 36.4%) and high-level resistance to streptomycin (30.0% vs 4.5%) in urban wild boars compared to rural wild boars (p < 0.05). E. faecalis was more frequently resistant to trimethoprim in urban than rural wild boars (33.3% vs 0.0%, p < 0.05). In isolates from urban origin, 55.6% of the likelihood of detecting antibiotic resistance depended only on the bacterial species, being more likely in the enterococci than in E. coli. These results suggest that urban wild boars may be more exposed to certain antibiotic-resistant bacteria or antibiotic resistance genes that they may acquire from the urban environment, although implications are uncertain.

© 2015 Deutsche Gesellschaft fü Säugetierkunde. The study of fluctuating asymmetry (FA) in living organisms has produced contradictory results over the past few decades of research. Though the protocol for measuring FA is firmly established, the sources of FA remain unclear in many cases. Our goal is to examine the relationship between FA and both the concentration of biomarkers of reactive oxygen species (ROS) and body condition in a medium-sized mammal, the European wild boar (Sus scrofa). Using a Partial Least Squares regression (PLSr), we found a positive significant relationship (Stone-Geisser test) between oxidative stress and FA but a negative relationship between oxidative stress and body condition. Our results suggest that FA can be used to assess the physiological costs associated with oxidative stress in mammals.

[This corrects the article DOI: 10.1371/journal.pone.0163971.].

Campylobacter infections are a public health concern and an increasingly common cause of food-borne zoonoses in the European Union. However, little is known about their spill-over from free-ranging livestock to sympatric wild ungulates, especially in regards to uncommon Campylobacter species. In this study, we aim to determine the prevalence of C. coli, C. jejuni and other C. spp. in game ungulates (wild boar Sus scrofa and Iberian ibex Capra pyrenaica) and free-ranging sympatric cattle in a National Game Reserve in NE Spain. Furthermore, we explore the extent to which Campylobacter species are shared among these co-habiting hosts. Faecal samples from Iberian ibex (n = 181) were negative for C. spp. By direct plating, two wild boars out of 150 were positive for C. coli (1.3%, 95% CI 0.16-4.73), and one was positive for C. jejuni (0.67%, 95% CI 0.02-3.66). The latter was predominant in cattle: 5.45% (n = 55, 95% CI 1.14-5.12), while C. coli was not isolated from this host. C. lanienae was the most frequent species in wild boar at 10% (95% CI 5.7-15.96), and one cow cohabiting with positive wild boars in the same canyon also carried C. lanienae. Four enrichment protocols (using Bolton or Preston broth combined with either mCCDA or CFA) were added for 172 samples (57 from wild boars, 55 cattle and 60 Iberian ibexes) to increase the number of isolates obtained allowing the detection of statistically significant differences. The prevalence of C. lanienae was statistically significantly higher in wild boar than in cattle (P < 0.01), but the prevalence of C. jejuni was higher in the latter (P = 0.045). These results suggest that wild boar and cattle carry their own predominant Campylobacter species, while Iberian ibex do not seem to play an important role in the epidemiology of Campylobacter. However, there is a potential spill-over of C. spp., and thus, further research is needed to elucidate the factors determining inter-species transmission.

Wildlife is increasingly abundant in urban environments, but little is known about the zoonotic pathogens carried by these populations. Urban wild boars are of particular concern because this species is well-known as a pathogen reservoir, and thus, we studied selected zoonotic pathogens in urban wild boars in Barcelona, Spain ( n = 41). Salmonella enterica was found in 5.00% (95% CI 0.61–16.91) and Campylobacter coli in 4.88% (95% CI 0.6–16.53) of the animals. E. coli O157:H7 and C. jejuni were not found. Other thermophilic Campylobacter were moderately prevalent (19.51%, 95% CI 8.82–34.87). Additionally, we screened for antimicrobial resistance in indicator bacteria: resistance was most frequent in Enterococcus faecium (95% of the isolates were resistant to at least one antimicrobial agent), followed by Enterococcus faecalis (50%) and Escherichia coli (10%). For the first time resistance to linezolid in bacteria carried by wildlife is reported. These findings pose a concern for public health, and thus, further research is needed on wildlife in urban environments.

Methicillin-resistant Staphylococcus aureus (MRSA) is a life-threatening pathogen in humans and its presence in animals is a public health concern. The aim of this study was to measure the prevalence of MRSA in free-living wild animals. Samples from red deer (n= 273), Iberian ibex (n= 212), Eurasian Griffon vulture (n= 40) and wild boar (n= 817) taken from different areas in Spain between June 2008 and November 2011 were analyzed. Characterization of the isolates was performed by spa typing, multi-locus sequence typing (MLST) and antimicrobial susceptibility testing. A low prevalence of MRSA was found with 13 isolates obtained from 12 animals (0.89%; 95% CI: 0.46-1.56). All MRSA sequence types belonged to ST398 (t011 and t1451) and ST1 (t127). Genotypes and antimicrobial susceptibility patterns (tetracycline resistance in ST398 and clindamycin-erythromycin-tetracycline resistance in ST1) suggest that the MRSA found probably originated in livestock (ST398) or humans (ST1). This is the first report of MRSA carriers in free-living wild animals in Europe. Although our data showed that MRSA prevalence is currently low, free-living wild animals might act as reservoir and represent a potential risk for human health., This work was partially supported by the Ministry of Science and Innovation within the Program of Interaction between wild animals and livestock (FAU200600011; FAU2008-00021) and by the Autonomous Community of Madrid, Spain (S0505/AGR-0265; S2009/AGR-1489). Eurasian Griffon vultures were captured within the Territorial Cooperation Programme Spain-France-Andorra (NECROPIR-EFA 130/09). E. Serrano was supported by the Beatriu de Pinós programme (BP-DGR 2011) of the Catalan Science and Technology System and N. Navarro-Gonzalez by the FPU programme of the Ministry of Education (Spain).

The genus Salmonella is found throughout the world and is a potential pathogen for most vertebrates. It is also the most common cause of food-borne illness in humans, and wildlife is an emerging source of food-borne disease in humans due to the consumption of game meat. Wild boar is one of the most abundant European game species and these wild swine are known to be carriers of zoonotic and food-borne pathogens such as Salmonella. Isolation of the pathogen, serotyping and molecular biology are necessary for elucidating epidemiological connections in multi-host populations. Although disease management at population level can be addressed using a number of different strategies, such management is difficult in free-living wildlife populations due to the lack of experience with the wildlife-livestock interface. Herein, we provide the results of a 4-year Salmonella survey in sympatric populations of wild boar and cattle in the Ports de Tortosa i Beseit National Game Reserve (NE Spain). We also evaluated the effects of two management strategies, cattle removal and increased wild boar harvesting (i.e. by hunting and trapping), on the prevalence of the Salmonella serovar community. The serovars Meleagridis and Anatum were found to be shared by cattle and wild boar, a finding that was confirmed by 100% DNA similarity patterns using pulse field gel electrophoresis. Cattle removal was more efficient than the culling of wild boar as a means of reducing the prevalence of shared serotypes, which underlines the role of cattle as a reservoir of Salmonella for wild boar. To our knowledge, this is the first attempt to manage Salmonella in the wild, and the results have implications for management.

Author(s): Navarro-Gonzalez, Nora; Jay-Russell, Michele T. | Abstract: Some species of birds form large flocks that forage on agricultural fields and frequently damage the crop, resulting in significant economic losses. Additionally, by defecating they potentially contaminate the crop and farm equipment with pathogens, which is of special concern for leafy greens that are consumed uncooked. There is a need to find an effective deterrent that is also environmentally friendly, and falconry is an ideal candidate for this purpose. To evaluate falconry as deterrent of nuisance birds in leafy greens field in northern California, we performed a set of trials on a control ranch and a treatment ranch of similar production and landscape characteristics. We identified avian species, counted individuals, and recorded flock size in daily surveys pre-treatment, during treatment (falconry), and post-treatment. Bird abundance was lower in spring than in fall, probably because many fields were fallow in spring. In both seasons, we observed a large daily variation in the bird abundance and their use of the fields in the surveys pre-treatment. In fall, the first trial was interfered by the activity of a falconer in an adjacent vineyard and the harvest of the grapes. Importantly, in the second trial, use of the field (e.g., foraging, etc.) decreased during five days of treatment and continued to be low for three days post-treatment, suggesting a “memory effect” after hazing by falconry. Interestingly, the third fall trial coincided with the end of the leafy greens growing season and showed that falconry successfully minimized use of the field by nuisance birds during peak activity. These results indicate that falconry is an effective measure to protect leafy green crops from fecal contamination and damage, but further research is needed at more farms in different regions, and the effect of using falconry in combination with other non-lethal bird abatement approaches (e.g., audio-visual deterrents) should also be evaluated.

Monitoring nutritional status of wildlife is key to understanding the population response to changes in food availability. Several direct and indirect methods have been proposed for such purposes, but faecal nitrogen (FN) is by far the most commonly used indirect non-invasive indicator of free-ranging ruminant diet quality. In this work, two methods were used to estimate the FN content in 291 pellet samples of Pyrenean chamois (Rupicapra pyrenaica pyrenaica) collected during a 2-year period (May 2009 to May 2011) in two chamois populations from the Catalan Pyrenees, Spain: the Dumas LECO analyzer and near-infrared reflectance spectroscopy (NIRS). In order to optimize further FN estimations we performed three different NIRS calibrations: (1) general, across year, season and populations; (2) population-specific; and (3) season-specific, i.e., different periods of vegetation growth. FN ranged from 1.66 % to 2.85 % (dry matter basis) and was higher in the main vegetation growing season than in winter. Concerning NIRS calibration, our study confirmed that FN of Pyrenean chamois can be accurately determined by NIRS, since the general calibration was accurate enough to predict FN. It is concluded that NIRS represents a non-destructive, emission-free and low-cost analytical technique which may reduce the use of conventional laborious methods for estimating FN in long-term wildlife monitoring.

International audience; The influence of landscape structure and host diet on parasite load of wildlife is still largely unknown. We studied a roe deer () population in a fragmented agricultural landscape in southern France to explore the relationship of gastrointestinal nematode load with spleen mass (to index immunocompetence), faecal nitrogen (to index diet quality), landscape structure and age of 33 hunt-harvested roe deer. Gastrointestinal worm counts were negatively related to faecal nitrogen and spleen mass, explaining respectively 24.7% and 9.2% of the observed variability in parasite load. Landscape structure did not appear to have a direct influence on gastrointestinal worm counts, but since animals from more open areas have a diet that is richer in nitrogen, its influence may be indirect. In conclusion, in the study area, the colonisation of the agricultural landscape does not seem to have increased the risk of gastrointestinal nematode parasitism for roe deer, possibly because access to high-quality food enhances immunocompetence.

The European wild rabbit Oryctolagus cuniculus has been recently reclassified as Near Threatened in its natural range in the Red List of Endangered Species by the IUCN, and a huge conservation effort is being undertaken in Spain for this keystone species. Restocking is a frequent measure for wild rabbit population reinforcements and it is also part of predator conservation programmes. However, it can have a negative influence on the resident rabbit population when it is not carried out carefully. In our work, using a model selection procedure based on a theoretic information approach, we analyze which factors favor the presence of sarcoptic mange in some wild rabbit populations in Catalonia (north-eastern Spain), as well as the trend of rabbit abundance in affected and non-affected hunting estates. Presence of mange depends on animal abundance and on restocking numbers (22.09%). From the mean rabbit abundance (30 rabbits hunted km � 2 ) and the mean restocking rate (18 rabbits km � 2 ), the probability of being affected increased in nearly 5% when the restocking rate increased in one unit. Rabbit abundance (2001–2007) depends on the presence of mange and on the effect of the year (23.86%), and clearly declined in the hunting estates with mange, whereas abundance is maintained in mange-free zones. Our results indicate that a sanitary control is necessary when restocking wild rabbit populations. Furthermore, restocking is shown to be a potential way of introducing pathogens to resident populations, especially under conditions of high density.

Perphenazine enanthate was used to allow adaptation to captivity in 11 Pyrenean chamois (Rupicapra pyrenaica). At the time of capture, all animals received 0.10 mg/kg of acepromazine maleate and 2.5 mg/kg of perphenazine enanthate intramuscularly. The effect was evaluated by means of three behaviors: alertness, defecation, and flight distance. The tranquilization and lack of fear of humans of all animals were determined and the usefulness of this long-acting tranquilizer for chamois adaptation to captivity was confirmed.

Wild ungulates have greatly increased in abundance and range throughout Europe. This new situation presents a concern for public health because many wild ungulates are known reservoirs of zoonotic pathogens.In this work, we tested for the presence of the zoonotic pathogen Escherichia coli O157:H7 in free-ranging livestock and sympatric wild boars (Sus scrofa) and Iberian ibex (Capra pyrenaica) in NE Spain from 2009 to 2011. In addition, antimicrobial resistance and virulence factors were assessed.In total, individual fecal samples were obtained from 117 hunter-harvested wild boars and 160 Iberian ibexes. Fifty-five samples were obtained from cattle (5 herds, 380 animals in total) and four from the only horse herd in the Natural Park 'Ports de Tortosa i Beseit' (32 animals). Fecal samples were processed according to the ISO 16.654:2001 protocol to obtain E. coli O157 based on immunomagnetic separation. In addition, a multiplex polymerase chain reaction (PCR) targeting nine virulence factors characteristic of human pathotypes was performed. The prevalence was compared between host species with Fisher's exact test.Four wild boars (3.41%, 95% CI = 0.94-8.52) and two Iberian ibexes (1.25%, 95% CI = 0.15-4.4) carried E. coli O157:H7, which was not found in livestock feces (n = 59, 95% CI = 0-8.94). All E. coli O157:H7 isolates were susceptible to all antimicrobial agents tested.These results indicate that when prevalence in co-habiting livestock is low, wild ungulates do not seem to play an important role as reservoirs of E. coli O157:H7.

The most widely used technique to assess helminth infection in both domestic and wild mammals is the faecal egg count (FEC). Most efforts to test the reliability of FEC as a proxy for parasite load are in small ruminant studies and limited work has evaluated the use of FEC in pigs. The aim of this study was to explore whether FEC is a reliable indicator of helminth load, and to evaluate the effects of sample storage on FEC accuracy in 59 wild boars. Though FEC was useful for assessing most helminth infections (e.g., Metastrongylus spp., Ascaris suum, Trichuris suis), stomach nematodes were often missed. The accuracy of FEC decreased over time, and thus it is recommended that samples be processed within 5 days of collection.

Antimicrobial resistance was assessed in indicator Escherichia coli isolates from free-ranging livestock and sympatric wild boar ( Sus scrofa ) and Iberian ibex ( Capra pyrenaica ) in a National Game Reserve in northeastern Spain. The frequency of antimicrobial resistance was low (0% to 7.9%). However, resistance to an extended-spectrum cephalosporin and fluoroquinolones was detected.

We determined whether different methods to isolate Campylobacter (including the ISO standard 10272:2006-1) affected the genotypes detectable from poultry, at three points during slaughter: caecal content, neck skin and meat. Carcasses from 28 independent flocks were thus sampled (subset A). In addition, ten neck skin samples from four flocks, ten caecal samples from ten different flocks and ten unrelated meat samples obtained from local supermarkets were collected (subset B). Campylobacter was isolated using eight different protocols: with and without enrichment using Bolton broth, Preston broth or Campyfood broth (CFB), followed by culture on either modified Charcoal Cefoperazone Deoxycholate Agar (mCCDA) or Campyfood agar (CFA). All obtained isolates were genotyped for flaA-SVR, and over half of the isolates were also typed by MLST. The strain richness, as a measure of number of detected fla-genotypes, obtained from subset A neck skin and caecal samples was higher than that of meat samples. In half of the cases, within a flock, at least one identical fla-genotype was obtained at all three slaughter stages, suggestive of autologous contamination of carcasses. Enrichment reduced the observed richness of isolates, while CFA plates increased richness compared to mCCDA plates, irrespective of inclusion of an enrichment step. Because the isolation protocol used influences both the yield and the fla-genotype richness obtained from poultry, this variable should be taken into account when different studies are being compared. Significance and Impact of Study The tracing of Campylobacter through the food chain remains important to control campylobacteriosis in humans. Our study points out that the isolation method used affects the genotypes obtained, and this should be considered as a variant when comparing the results of surveillance studies.

SummarySalmonella enterica is a zoonotic agent of worldwide importance found in a widerange of wild hosts. However, its prevalence in many popular game species hasnever been assessed. Iberian ibex (Capra pyrenaica) is the main game caprinae ofthe Iberian Peninsula and around two thousand individuals are hunted every yearfor trophy or for home consumption. In this work, 313 Iberian ibexes from thePorts de Tortosa i Beseit National Game Reserve (NE Spain) were tested for Sal-monella enterica in faeces, and anti microbial susceptibility was determined. Theexact location of shooting or capture was recorded with a GPS device to study thelinks of Salmonella infection with cattle presence and human proximity. Addi-tionally, samples were taken from cattle grazing inside this reserve (n = 73). Onlythree Iberian ibexes (0.96%, 95% CI 0.2–2.8) were positive to Salmonella (sero-type Enteritidis, Bardo and 35:r:z35), while prevalence was moderate in cattle:21.92% (95% CI 13.10–33.14, serotype Meleagridis, Anatum, Kedougou andOthmarschen). All isolates were susceptible to the anti microbial agents tested.Moreover, a case of fatal septicaemic salmonellosis in an 11-year-old male Iberianibex is described where Salmonella enterica serotype Enteritidis was isolated fromthe lung, liver and spleen samples. The low prevalence of Salmonella in Iberianibex and the lack of shared serotypes suggest no association to cattle. Despite this,game meat aimed for human consumption should be examined, and it is stronglyrecommended that hunters and game keepers manipulate animals and carcassesunder maximal hygienic conditions to avoid environmental contamination andhuman contagion.IntroductionSalmonellosis continues to be the second most frequentzoonoses in the European Union (EFSA, 2012). However,many gaps exist in our knowledge about its presence anddistribution in wildlife, which has been stated to be a com-mon reservoir. Particularly, game species may play a moreimportant role in its epidemiology, displaying an increasedrisk for public health owing to (i) its meat being consumedby humans and (ii) being abundant species potentiallyinvolved in environmental contamination (e.g. watersources, recreational waters or pastures). In fact, diseasetransmission between sympatric wildlife and livestock canbe of particular concern in relation to wild ungulates,which frequently share habitat resources with domesticlivestock (Boehm et al., 2007).The Iberian ibex is among the species about which thereis a lack of information. It is the main game caprinae of the

Wild boar is a recognized reservoir of bovine tuberculosis (TB) in the Mediterranean ecosystems, but information is scarce outside of hotspots in southern Spain. We describe the first high-prevalence focus of TB in a non-managed wild boar population in northern Spain and the result of eight years of TB management. Measures implemented for disease control included the control of the local wild boar population through culling and stamping out of a sympatric infected cattle herd. Post-mortem inspection for detection of tuberculosis-like lesions as well as cultures from selected head and cervical lymph nodes was done in 745 wild boar, 355 Iberian ibexes and five cattle between 2004 and 2012. The seasonal prevalence of TB reached 70% amongst adult wild boar and ten different spoligotypes and 13 MIRU-VNTR profiles were detected, although more than half of the isolates were included in the same clonal complex. Only 11% of infected boars had generalized lesions. None of the ibexes were affected, supporting their irrelevance in the epidemiology of TB. An infected cattle herd grazed the zone where 168 of the 197 infected boars were harvested. Cattle removal and wild boar culling together contributed to a decrease in TB prevalence. The need for holistic, sustained over time, intensive and adapted TB control strategies taking into account the multi-host nature of the disease is highlighted. The potential risk for tuberculosis emergence in wildlife scenarios where the risk is assumed to be low should be addressed.

Salmonella is distributed worldwide and is a pathogen of economic and public health importance. As a multi-host pathogen with a long environmental persistence, it is a suitable model for the study of wildlife-livestock interactions. In this work, we aim to explore the spill-over of Salmonella between free-ranging wild boar and livestock in a protected natural area in NE Spain and the presence of antimicrobial resistance. Salmonella prevalence, serotypes and diversity were compared between wild boars, sympatric cattle and wild boars from cattle-free areas. The effect of age, sex, cattle presence and cattle herd size on Salmonella probability of infection in wild boars was explored by means of Generalized Linear Models and a model selection based on the Akaike’s Information Criterion. Prevalence was higher in wild boars co-habiting with cattle (35.67%, CI 95% 28.19–43.70) than in wild boar from cattle-free areas (17.54%, CI 95% 8.74–29.91). Probability of a wild boar being a Salmonella carrier increased with cattle herd size but decreased with the host age. Serotypes Meleagridis, Anatum and Othmarschen were isolated concurrently from cattle and sympatric wild boars. Apart from serotypes shared with cattle, wild boars appear to have their own serotypes, which are also found in wild boars from cattle-free areas (Enteritidis, Mikawasima, 4:b:- and 35:r:z35). Serotype richness (diversity) was higher in wild boars co-habiting with cattle, but evenness was not altered by the introduction of serotypes from cattle. The finding of a S. Mbandaka strain resistant to sulfamethoxazole, streptomycin and chloramphenicol and a S. Enteritidis strain resistant to ciprofloxacin and nalidixic acid in wild boars is cause for public health concern. This work has been partially funded by the Spanish Ministry of Science and Innovation within the Program of Interaction between wild animals and livestock (FAU2006-00011 and FAU2008-00021). N. Navarro-Gonzalez was supported by the FPU programme of the Ministerio de Educación (MEC, Spain) and E. Serrano by the Beatriu de Pinós programme (BP-DGR 2011) of the Catalan Science and Technology System (Spain). Phage Typing was performed in the Centro Nacional de Microbiología, Instituto de Salud Carlos III, Madrid, Spain) within a collaborative project funded by the Comunidad de Madrid (Vigilancia Sanitaria, S2009/AGR-1489). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.